摘要
目的利用大肠杆菌系统表达乙型肝炎病毒表面抗原″ａ″决定簇。方法和结果采用ＰＣＲ技术，以ＨＢＶａｙｗ２亚型为模板，扩增ＨＢｓＡｇ″ａ″决定簇（１２３－１４７ａａ）基因片段（８５ｂｐ），经ＨｉｎｄⅢ酶切后，克隆到质粒载体ＰＭＴ－ｈＩＬ３中，成功构建了ＰＭＴ－ａ重组质粒。结论经４２℃诱导表达，表达产物占菌体总蛋白２８％，分子量１６０００，具有″ａ″决定簇抗原性。
Expression and characterization of HBsAg 'a' determinant in E. Coli. Methods and Results A size of 85bp fragment. which encodes the entire HBsAg 'a' determinant from 123 to 147 amino acid sites of HBsAg, was amplified with HBV ayw2 subtype as template by using PCR technique. After digestion with restriction enzyme Hind Ⅲ, the PCR product was cloned into expression vector PMT-hIL3. Cohclusions During heat induction at 42℃, the resulting recombinant plasmid PMT-a could overproduce a 16kD fusion protein in E. Coli at a level of 28% of total cellular proteins. ELISA analysis showed that the fusion protein conserved the antigenicity of 'a' determinant.
出处
《中华肝脏病杂志》
CAS
CSCD
1996年第3期151-153,共3页
Chinese Journal of Hepatology
基金
国家自然科学基金
霍英东青年教师研究基金
